The Globalization of Pharmaceutics Education Network (GPEN ...



TABLE OF CONTENTS

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SECTION PAGE

Welcome.....................................................................2-4

Acknowledgments.....................................................5-7

Participants

Keynote Speaker………………………..…...8

University/Industry Attendees………………9-15

Program

Wednesday, November 6, 2002....................15-18

Thursday, November 7, 2002……………...19-21

Friday, November 8, 2002…………………22

Presented Posters…………………………..23-31

Podia Abstracts

Podia Assignments………………...............32-33

Podia Abstracts…………………………....34-60

Poster Abstracts

Poster Assignments……………...…….….61-65

Poster Abstracts…………………………..66-136

Short Courses

Scientific Short Courses Schedule……….137-144

Notes

Contacts……….………………………….145-146

Meeting Notes............................................147-150

WELCOME

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The Globalization of Pharmaceutics Education Network (GPEN), was initiated by the University of Kansas in early 1996 to establish an international network of pharmaceutics programs and to facilitate the exchange between faculty and graduate students among participating programs.  Since its founding GPEN has biennially held major conferences which serve as a premier meeting ground and forum for pharmaceutical sciences research and education, providing its 31 member institutions with an arena for graduate students to share new ideas, hear international leaders in various research specialties, and network with current and future pharmaceutical scientists from industry and academia.  These conferences draw pharmaceutical scientists from many parts of the United States as well as from several countries around the world. To date, GPEN meetings have been held at The University of Kansas, Lawrence, KS USA (October 24-26, 1996), The Swiss Federal Institute of Technology (ETH) of Zürich, Zürich, Switzerland (September 8-10, 1998), and Uppsala University, Uppsala, Sweden (September 13-15, 2000).

WELCOME

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Pharmacy was first taught at the University of Michigan in 1868 in the College of Literature, Science and the Arts. It was in 1876 that a School of Pharmacy was established, the first in a state university in the United States. In 2001, the College celebrated its 125th anniversary, commemorating 125 glorious years of education, research and service since the founding of the College as a separate department within the University.

Although the College offers several academic programs, for decades the College’s Graduate Program in Pharmaceutics has been ranked among the best in the world. With a total enrollment of approximately 80 students, Ph. D. scholars at the College have personal access to some of the world’s most eminent scientists and educators. This is an ideal environment to learn and research. Our scholars have access to all the facilities offered by a huge university, yet enjoy the advantages of belonging to a small, close-knit group.

We welcome all the GPEN attendees to our College and our department. We invite you to share our world for the time that you are here, and experience our program as we know it – an inspiring one.

GPEN Conference Schedule

Tues., Nov.5 Wed., Nov 6 Thurs., Nov. 7 Fri., Nov 8 Sat., Nov. 9

|8:00 AM |Arrival |Welcome/ Student Podia |Student Podia |Scientific Short Courses|Departure |

|9:00 AM | | | | | |

|10:00 AM | | | | | |

|11:00 AM | | | | | |

|12:00 PM | |Lunch and |Lunch and |Lunch | |

| | |Poster Session |Poster Session | | |

|1:00 PM | | | |Scientific Short Coures | |

| | | | | | |

| | | | | | |

| | | | | | |

| | | | | | |

| | | | |End of meeting | |

|2:00 PM | |Student Podia |Student Podia | | |

|3:00 PM |Pfizer Tour | | | | |

|4:00 PM | | | | | |

| | | |Break | | |

|5:00 PM |Reception |Break | | | |

|6:00 PM | | | | | |

|7:00 PM | |Dinner |Banquet/ Keynote Address | | |

| | | | | | |

ACKNOWLEDGMENTS

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Academic Sponsors

University of Michigan College of Pharmacy

University of Michigan Horace Rackham School of Graduate Studies

American Association of Pharmaceutical Scientists

Special Recognition

The 2002 GPEN Organizing Committee would like to thank several individuals who made important contributions to this year’s meeting.

• Professors Gordon L. Amidon and Ronald T. Borchardt for their guidance, help, and support

• Dr. Peter Green and Dr. Catherine Knupp for their guidance and support in organizing the Pfizer tour, and for assisting in guest speaker selection

• Dr. Arthur Franke for his presentation

• Pharmaceutical Sciences faculty

• Professor Wang for the support of the Pharmaceutical Engineering program

• Poster and podia presenters

• Industrial representatives

ACKNOWLEDGMENTS

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• Graduate students, post-doctoral fellows, and researchers from the Pharmaceutical Sciences Department at the University of Michigan School of Pharmacy

• Nancy Helm at the University of Kansas and Terri Jasin at the University of Michigan for their administrative support.

• Florence Gerber at Pfizer for organizing the departmental tours

• Jim Gannon and the University of Michigan Copy Center staff for their assistance in producing GPEN program booklets

• Karen Samas and the Michigan League staff for their help in planning of events

GPEN 2002 Organizing Committee

Faculty Advisors

Professor Gordon Amidon

Professor Ronald Borchardt

Meeting Chairs

Chris Landowski

Phil Lorenzi

Maureen Connell

ACKNOWLEDGMENTS

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Financial Committee

Scott Ocheltree(Chair)

Sachin Mittal

Chris Landowski

Program Committee

Sarita Naik (Chair)

Balvinder Vig

Amy Ding

Chengi Cui

Sujatha Menon

Chris Landowski

Social/FoodCommittee

Maureen Connell(Chair)

Sachin Mittal

Kiarri Kershaw

Theresa Nguyen

Yongtao Li

Insook Kim

Ying Li

Audio Visual Committee

Filippos Kesisoglou

Yongtao Li

Keynote Speaker: Dr. Arthur Franke

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Arthur Franke, Ph.D., is Sr. Research Director in Molecular Sciences and Technologies at Pfizer Global Research and Development Laboratories in Ann Arbor, MI. In this role, Dr. Franke currently oversees drug discovery programs in Bioinformatics, Expression Profiling, Proteomics, and Biomarkers. Dr. Franke has been actively exploring innovative applications of molecular sciences to pharmaceutical research for the past 20 years.

Dr. Franke obtained his undergraduate and doctoral degrees in Microbiology from the University of Michigan, Ann Arbor. After completing postdoctoral research at Genentech, Inc., Dr. Franke began his career at Pfizer in Groton, CT as a Research Scientist. His career at Pfizer has been marked by strong scientific contributions beginning with genetic engineering research that generated proprietary expression cultures used to develop a commercial process for Pfizer's first product derived from recombinant DNA technology.

Dr. Franke has a long record of strong scientific leadership in the application of genetic technologies to drug discovery processes. Current research activities focus on building high quality target portfolios in psychotherapeutics, cancer, anti-bacterials and other therapeutic areas. Dr. Franke has been recognized as a key contributor to several external collaborations in Pfizer’s global network of discovery technologies.

As the GPEN 2002 guest speaker, Dr. Franke will be presenting his keynote address, entitled "The Application of Genomics/Proteomics to Drug Discovery.”

PARTICIPANTS

Industrial Attendees

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Pfizer

|Linda Hitchingham |David Pipkorn |

|Yung-Chi Lee |Chris Seadeek |

|Shouchin Man | |

Becton-Dickenson

|P Green | |

Pharmacia

|P Burton |Hong Qi |

|Richard Hwang | |

|Clayton Jacobsen | |

Esperion

| | |

|R Newton | |

Lilly

| | |

|Amin Khan |Martha Kral |

Forrest Laboratories

| | |

|Andreas Grill |Shashank Mahashabde |

PARTICIPANTS

University Attendees

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Swiss Federal Institute of Technology-Zürich (ETHZ)

| | |

|Natalia Blarer |Vera Luginbühl |

|Samantha Jilek |Hans Merkle (F) |

|Annette Koch | |

Hebrew University

| |Abraham Rubinstein (F) |

|Kareem Azab |Boris Vaisman |

|Simon Benita (F) | |

Kyoto University

|Naoki Kobayashi |Yoshinobu Takakura (F) |

|Masato Maruyama |Kei Yasuda |

Leiden/Amsterdam Center for Drug Research (LACDR)

| | |

|Maytal Bivas-Benita |H Junginger (F) |

|Gerrit Borchard (F) | |

Phillips University

| | |

|Dagmar Fischer (F) |Christine Oster |

|Thomas Merdan | |

Royal Danish School of Pharmacy

| | |

|Andre Eriksson |Vibeke Hougaard Sunesen |

|Sven Frokjaer (F) | |

University of Basel

|Georgios Imanidis (F) |Marc Sutter |

*All participants listed are graduate students, with the exception of Faculty, denoted by (F).

PARTICIPANTS

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University of Kansas

|Meagan Anderson |Vikram Sadineni |

|Kenneth Audus (F) |Christian Schöneich (F) |

|Ronald Borchardt (F) |Teruna Siahaan (F) |

|Arvind Chappa |Sutthilug Sotthivirat |

|Josh Cooper |Valentino Stella (F) |

|Ajit Joseph D’Souza |Carol Stotz |

|Nathan Lacher |Fei Tian |

|Susan Lunte (F) |Jiaher Tian |

|R Middaugh (F) |Liz Topp (F) |

|Eric Munson (F) |Amber Young |

|Stephanie Pasas | |

University of Michigan

| | |

|Gordon Amidon (F) |Phil Lorenzi |

|Pen-Chung Chen |Elizabeth Matthew |

|Chengji Cui |Sujatha Menon |

|Karen Dehring |Len Middleton (F) |

|Amy Ding |Sachin Mittal |

|Gordon Flynn (F) |Cheol Moon |

|Jichao Kang |Sarita Naik |

|Richard Keep (F) |Theresa Nguyen |

|Ron Kelly |Scott Ocheltree |

|Kiarri Kershaw |Gus Rosania (F) |

|Filippos Kesisoglou |Steven Schwendeman (F) |

|Insook Kim |David Smith (F) |

|Longsheng Lai |Barbara Spong |

|Chris Landowski |Ethan Stier |

|Lei Li |Balvinder Vig |

|Ying Li |Michele Van de Walle (F) |

|Yongtao Li |Jennifer VanRoeyen |

|Jun Feng Liang |Henry Wang (F) |

|Zheng Lu |Victor Yang (F) |

|Kyung-Dall Lee (F) |Philip Zocharski |

PARTICIPANTS

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|University of Tokyo | |

| | |

|Maki Hasegawa |Hiroshi Suzuki (F) |

|Kazuya Maeda | |

University of Utah

| | |

|Padmanabh Chivukula |W. Higuchi (F) |

|David Day |Jindrich Kopeček (F) |

|Hui Ding |Chunyu Xu |

|Matthew Fiddler |Guang Yan |

|Ram Goteti | |

University of Nebraska

| | |

|Corbin Bachmeier |Dr. Miller (F) |

|Haiqing Dai |Jayanth Panyam |

|Sinjan De |Swayam Prabha |

|William Elmquist (F) |Sanjeeb Sahoo |

Uppsala University

| | |

|Per Artursson (F) |Niclas Rydell |

|Niclas Petri |Lena Strindelius |

|Eva Ragnarsson | |

Victorian College of Pharmacy

| | |

|Susan Charman (F) |Richard Prankerd (F) |

|Danielle McLennan |Alicia Segrave |

|Joseph Nicolazzo | |

University of Colorado

| | |

|Derrick Katayama |Mark Manning (F) |

|Ng Lawrence (F) |Huiyu Zhou |

PARTICIPANTS

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University of Connecticut

| | |

|Harminder Bajaj |Vikas Sharma |

|Bakul Bhatnager |Charlie(Xiaolin) Tang |

|Michael Pikal (F) | |

Saarland University

| | |

|Carsten Ehrhardt |Ulrich Schafer (F) |

|Claus-Michael Lehr (F) | |

Utrecht University

| | |

|Wim Hennink (F) |Karin Wensink |

|Bernard Mets | |

Hacettepe University

| | |

|Hakan Eroğlu |Erem Memişoğlu |

Kanazawa University

| | |

|Akihiro Inano (F) |Dr. Tsuji (F) |

|Akiko Takamoto (F) | |

Katholieke University of Leuven

| | |

|Remmy Agu |Uche Dominic Obimah |

|Patrick Augustijns (F) |Karel Six |

|Annelies Derycke | |

PARTICIPANTS

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University of Florida

| | |

|Vikram Arya |Qi Liu |

|Whocely Victor de Castro |Irene Rinas |

|Derendorf Hartmut (F) |Edgar Schuck |

|Ariya Khunvichai |Virna Schuck |

|Julia Kurz |Immo Zdrojewski |

|Ping Liu | |

University of Kentucky

| | |

|Bradley Anderson (F) |Laura Land |

|Zhengrong Cui |Russ Mumper (F) |

|Tyler R. DeGraw |Moses Oyewumi |

University of Maryland

| | |

|Hamid Ghandehari (F) |Zak Megeed |

University of North Carolina

|David Bourdet |Keith Hoffmaster |

|K. Brouwer (F) |Dhiren Thakker (F) |

University of Otago

|Melissa Copland |Ian Tucker (F) |

|Natalie Medlicott (F) |Karen White |

University of Queensland

|Yiu-Ngok Chan |M Roberts (F) |

|Ross McGeary (F) |Istvan Toth (F) |

|Benjamin Ross | |

PARTICIPANTS

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University of Southern California

|Melina Bayramyan |Ashutosh Kulkarni |

|Michael Bolger |Vincent Lee (F) |

|Chun Chu |Jennifer Links |

|Hovik Gukasyan |Jeremy Links |

|Ian Haworth (F) |Jing (Crystal) Wang |

University of Kuopio

|Sari Peltola |Arto Urtti (F) |

|Marjukka Suhonen (F) | |

University of Geneva

|R Guy | |

Wednesday, November 6, 2002

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7:00 AM Breakfast—Ballroom

7:00-8:00 AM Registration—Ballroom

8:00 AM WELCOME—Ballroom

8:20 AM Development of a Bootstrap Procedure for Resampling Pharmacokinetic Data. Arya, V.1 Agarwal, A.1 Nunan, E.2 Santos, T.2 Hochhaus, G.1, 1University of Florida, Gainsville, 2Universidade Federal De Minas Gerias, Brazil

8:40 AM The Role of the Lymphatics in the Dose-Dependent Pharmacokinetics of Leukemia Inhibitory Factor in Sheep. Segrave, AM., Porter CJH, Charman SA. Monash University, Melbourne, Australia.

9:00 AM A Novel In Situ Technique for Direct Determination of First Pass Metabolism and Bi-directional Transport in the Rat Ileum. DeGraw, R., Anderson, B. University of Kentucky, KY

9:20 AM Human Alveolar Epithelial Cell Monolayers in Primary Culture as an In Vitro Model for Drug Absorption Studies. Ehrhardt C. Schaefer, UF., Lehr CM. Saarland University, Saarbruecken, Germany

9:40 AM Membrane Properties Affecting Drug Permeability. Sutter, M., Fiechter, T., Imanidis, G. University of Basel, Switzerland

Wednesday, November 6, 2002

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10:00 AM REFRESHMENTS

10:20 AM Quantitative Release of Active Lysozyme from Dextran Based Hydrogels. Wensink, KDF1.,Jiskoot, W.1, Verrijk, R.2, Hennink, WE.1, 1 Utrecht University, 2 Octopus Technologies b.v, Leiden

10:40 AM Selectivity of MTX-ICAM-1-peptide Conjugate Compared to MTX. Anderson, ME., Yakovleva T., Jean-Farve DJ., Siahaan TJ., The University of Kansas, KS.

11:00 AM New Boronated, Cationized Acrylamide Co-Polymers: Synthesis and Mucosal Attachment Properties. Azab, K., Srebnik, M., Rubinstein, A., The Hebrew University of Jerusalem, Israel.

11:20 AM Incorporation of Isoniazid into Poly(L-Lactide) Microspheres Using a Prodrug Approach. Zhou, H, Biggs, DL., Claffey, DJ., Ruth, JA.,Randolph, TW., Ng, K., Manning, MC., University of Colorado Health Sciences Center, CO.

11:40 AM LUNCH—Michigan, Vandenberg, Hussey Rooms

12:30-2:00 PM POSTER SESSION—Concourse

2:00 PM Liposomal Delivery of Antigen to Human Dendritic Cells. White, KL., Copland, MJ., Baird, MA., Hook, S., Davies, NM., Rades, T. University of Otago, New Zealand

Wednesday, November 6, 2002

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2:20 PM Phagocytosis of Biodegradable Microparticles to Induce Maturation and Activation of Dendritic Cells. Jilek, S., Merkle, HP., Walter, E. ETH Zürich, Switzerland

2:40 PM Flagellin from Salmonella Enteritidis Induce Strong Immune Responses after Oral and Nasal Vaccination in C3H/HeJ Mice. Strindelius, L., Sjöholm, I. Uppsala University, Sweden

3:00 PM Pulmonary DNA Vaccination Against TB using Chitosan Nanoparticles. Bivas-Benita, M., Geluk, A., van Meijgaarden, KE., van der Borch, F., Ottenhoff, THM., Junginger, HE., Borchardt, G. Leiden University, The Netherlands

3:20 PM Synthesis and Immunological Evaluation of Vaccines Containing Multiple Copies of Peptide Epitopes. Ross,BP.1, McGeary RP.1, Horváth, A.1, Olive, C.2, Good, M.2, Toth, I1. 1The University of Queensland, Australia; 2CRC for Vaccine Technology, Brisbane, Australia.

4-7:00PM BREAK

7:00 PM DINNER—Ballroom

8:00-9:30 PM ENTERTAINMENT—“The Friars”

Thursday, November 7, 2002

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7:00 AM Breakfast—Ballroom

8:15 AM OPENING REMARKS —Ballroom

8:20 AM Transferrin-Mediated Targeting of AlPcS4 Encapsulated in Sterically Stabilized Liposomes. Derycke, AS., de Witte, PA. KU Leuven, Belgium.

8:40 AM Design of a Lymphoma Targeted Drug Delivery System: Selection of CD21 Recognizable Epitopes with Phage Display. Ding, H.1, Prodinger, WM.2, Kopeček, J.1, 1University of Utah, UT; 2University of Innsbruck, Austria.

9:00 AM Preparation and Characterization of Progesterone-Loaded Amphiphilic (-Cyclodextrin Nanospheres. Memişoğlu, E., Bochot, A., Duchene, D., Hincal., A. Hacettepe University, Turkey; Université Paris-Sud, France.

9:20 AM Construction and Characterization of a Tissue Plasminogen Activator Mutant for Potential Use in Thrombolytic Therapy with Reduced Bleeding Risk. Naik, SS., Song H., Liang, JF., Yang, VC. The University of Michigan, MI.

9:40 AM REFRESHMENTS

10:00 AM Kinetics of Cold Denaturation in Viscous Systems and Protein Stability During Freeze Drying. Tang, X., Pikal, MJ. University of Connecticut, CT.

Thursday, November 7, 2002

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10:20 AM A New Poly(ethyleneimine)-graft Poly(ethylene glycol) Block Copolymer for Nucleic Acid Delivery In Vivo. Merdan, T., Kunath, K., Petersen, H., Voigt,K., Kissel, T. Philipps University, Marburg, Germany

10:40 AM Plasmid DNA Uptake and Subsequent Induction of Inflammatory Cytokines in Cultured Mouse Macrophages. Yasuda, K., Kawano, H., Takakura, Y. Kyoto University, Japan

11:00 AM Genetically Engineered Silk-Elastinlike Protein Polymers for Controlled Gene Delivery. Megeed, Z.1, Cappello, J.2, Ghandehari, H.1, 1University of Maryland, MD; 2Protein Polymer Technologies, Inc., San Diego, CA.

11:20 AM LUNCH—Michigan,Vandenberg, Hussey Rooms

12:30-2:00 PM POSTER SESSION—Concourse

2:00 PM Functional Involvement of Organic Anion Transporters in the Renal Uptake of Anionic Compounds and Nucleoside Derivatives. Hasegawa, M.1, Kusuhara, H.1,2, Endou, H.3, Sugiyama, Y.1,2, 1University of Tokyo, Japan; 2CREST, JST; 3Kyorin University

2:20 PM Refinement of a Computer Model of hPepT1 Structure-Function. Links, JLS., Haworth, IS, Lee, VHL. University of Southern California, CA.

Thursday, November 7, 2002

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2:40 PM Interactions of Chlorambucil and Melphalan with Drug Efflux Transporters in Brain Endothelial Cells. Bachmeier, CJ., Miller, DW. University of Nebraska Medical Center, NE.

3:00 PM Thymine Dipeptide Prodrugs:Aqueous Stability and In Vitro Affinity to the Human Di/Tripeptide Transporter, HPEPT1. Eriksson, AH., Steffansen, B. The Royal Danish School of Pharmacy, Denmark.

3:20 PM Studies on Functional Sites of Organic Cation/Carnitine Transporter OCTN2 (SLC22A5) Analyzed by Mutant Protein Found in Japanese Population. Inano, A., Ohashi, R., Tamai, I., Sai, Y, Tsuji, A. Kanasawa University, Japan

4:00-7:00 PM BREAK

7:00 PM BANQUET—Ballroom

KEYNOTE ADDRESS: "The Application of Genomics/Proteomics to Drug Discovery", Dr. Arthur Franke, Head of Genomics and Proteomics at Pfizer (Ann Arbor)

Friday, November 8, 2002 [pic]

7:00 AM Breakfast—Ballroom

8:00-12:15 PM Short Course 1 : Modern Molecular Biopharmaceutics: Transporters—Hussey Room (2nd floor)

8:00-5:00 PM Short Course 2: Designing Drugs with Optimal in vivo Activity Following Oral Administration—Michigan Room (2nd floor)

8:00-4:10 PM Short Course 3: Stability of Peptides, Proteins and Nucleotides—Vandenberg Room (2nd Floor)

8:00-12:15PM Short Course 4: Business Entrepreneurship—Room D (3rd Floor)

8:00-4:45 PM Short Course 5: Dermal and Transdermal Delivery—Koessler Room (3rd Floor)

12:30-4:00 PM Short Course 6: Barriers to CNS Drug Delivery / Transporters—Hussey Room (2nd Floor)

12:15-4:00 PM Short Course 7: Mucosal Vaccination—Room D (3rd Floor)

5:00 PM Conclusion of meeting

• Lunch will be served in the Ballroom from 11:30- 1:00 PM

• POSTER PRESENTATIONS

November 6-7, 2002

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Vaccine Delivery

Biodegradable Microspheres as Delivery System for Synthetic Antigens. Blarer, N., Merkle, HP., Gander B. ETH Zϋrich, Switzerland

A Method for the Incorporation of Ovalbumin into ISCOMs Prepared by the Hydration Method. Copland, MJ., Konnings, S., Davies, NM., Rades, T. University of Otago, New Zealand

Oral Vaccination against Diphtheria with Starch Micro-Particles as Adjuvant. Rydell, N., Sjohölm, I. Uppsala University, Sweden

Transporters

The Role of P-Glycoprotein in the CNS Distribution of a Novel Antitumor Agent, STI-571 (GLEEVEC). Dai, HQ., Shaik, N., Lemaire, M., Marbach, P., Elmquist, W. University of Nebraska Medical Center, NE

Localization or the Canalicular Transport Proteins MDR1 and MRP2 in Human Hepatocytes Cultured in a Sandwich Configuration. Hoffmaster, KA., Chandra, P., LeCluyse, EL., Brouwer, KLR. University of North Carolina, NC

Biophysical Evidence for His-57 as a Proton-binding

Site in the Mammalian Intestinal Dipeptide Transporter hPEPT1. Kulkarni, AA. 1, Uchiyama, T. 1, Loo, DDF. 2, Davies, DL. 1, Lee, VHL1. 1University of Southern California; 2UCLA School of Medicine, CA

Comparative Functional Analysis of Human OATP2 and OATP8. Maeda, K., Sugiyama, Y. University of Tokyo, Japan

POSTER PRESENTATIONS [pic]

Expression and Function of the Peptide Transporter, PHT1, in the Human Retinal Pigment Epithelium. Ocheltree, SM., Keep, RF., Shen, H., Hughes, BA., Smith, DE. University of Michigan, MI

Autoradiographic Detection of [3H]D- and [3H]L- Aspartate Binding to Glutamate Transporters in Frozen Sections of Rat Brain. Takamoto,A., Balcar, VJ., Yukio, Y. Kanazawa University, Japan.

Characterization of Efflux Transporters of the Human Trophoblast. Young, AM., Audus, KL. The University of Kansas, KS

Drug Delivery

Computer Modeling of Nucleic Acids. Bayramyan, MZ., Chambers, EJ., Price, EA., Haworth, IS. University of Southern California, CA.

Liposaccharide Delivery System for Peptide IT-100, LHRH and Conotoxin MII. Chan, Y.1, Blanchfield, J. 1, Rhee, H. 1, Alewood, P. 1, Adams, D. 1, Good, M. 2, Toth, I 1. University of Queensland; Queesnland Insitute for Medical Research, Australia.

Lidocaine-Releasing Microsphere-Gel System for In Vivo Localized Anesthetic Efficacy. Chen, PC. 1, Park, YJ. 1, Bartlett, R. 1, Kohane, D. 2, Chang, LC. 3, Langer, R. 2, Yang, VC1. 1The University of Michigan, MI; 2Massachusetts Institute of Technology, MA; 3Medical Defense University, Taiwan

Design of Interpenetrating Network Hydrogels for Colon-Specific Drug Delivery. Chivukula, P., Wang, D., Kopečkovà, Kopeček, J. University of Utah, UT

POSTER PRESENTATIONS

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Transmucosal Delivery of Calcitonin in Rabbits using Mucoadhesive Thin-Film Composites. Cui, Z., Mumper, RJ. University of Kentucky, KY

Polymeric Prodrug: Enhanced Release due to Intramolecular Ring Closure. D’Souza, AM., Topp, EM. The University of Kansas, KS

In Vitro Release Characteristics of Dexamethasone Sodium Phosphate from Chitosan Films. Eroğlu, H., Öner, L. Hacettepe University, Turkey

Identification and Characterization of a Prodrug Activating Enzyme: BPHL (Biphenyl Hydrolase-Like Hydrolyses Valacyclovir). Kim, I. 1, Chu, X. 2, Provoda, C. 1, Lee KD. 1, Amidon, GL1. 1The University of Michigan, MI; 2Merck and Co, NJ

The Use of a Transduction Peptide as a Possible Means for Protein Encapsulation. Li, YT, Liang, JF., Yang, VC. The University of Michigan, MI.

Local Release Kinetics of IGF I Loaded Microparticles Determine the Mechanism and Extent of Bone Repair. Luginbühl, V., Merkle, HP., Meinel L. ETH Zürich

Gadolinium Hexanedione Nanoparticles Engineered for Potential Application in Neutron Capture Therapy of Tumors. Oyewumi, MO., Mumper, RJ. University of Kentucky, KY.

Dynamics of Endocytosis and Exocytosis of PLGA Nanoparticles. Panyam, J., Labhasetwar, V. University of Nebraska Medical Center, NE.

Skin Penetration Pathways of FITC Insulin from Ethosomes. Vaisman B., Touitou E. The Hebrew University of Jerusalem, Israel

POSTER PRESENTATIONS

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Protein-Based Triblock Copolymers as Potential Drug Carriers. Xu, C., Kopeček. University of Utah, UT

Gene Delivery

Viral Gene Delivery to Migrating/ Wounded Intestinal Epithelial Cells. Kesisoglou, F., Schmiedlin-Ren, P., Chamberlain, JR., Roessler, BJ., Fleisher, D., Zimmermann, E. The University of Michigan, MI.

Characterization of Hepatic Delivery of Macromolecules with Large Volume Injection and its Therapeutic Application. Kobayashi, N., Kawase A., Takakura, Y. Kyoto University, Japan

Analysis of Secretion Polarity after Gene Delivery to Epithelial Cells. Maruyama, M., Watanabe, Y., Takakura, Y. Kyoto University, Japan

Cationic Nanoparticles Consisting of Polyethyleneimine and PLGA as Parenteral Carriers for DNA Vaccination. Oster, CG., Kissel, T. Philipps University, Germany

A Non-Viral Gene Delivery Formulation Stabilizes Reporter Gene Production In Vivo. Ragnarsson, EGE., Artursson, P., Köping-Höggård, M., Regnström. Uppsala University, Sweden

Biopharmaceutics

Complex Mechanism of Absorptive H-2 Antagonist Transport Across Caco-2 Cell Monolayers. Bourdet, DL., Hong, S., Ng, C., Brouwer, KLR., Thakker, D. University of North Carolina, NC

POSTER PRESENTATIONS

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Cholecystokinin Metabolism in Canine Buccal Mucosa. Day, DE. 1, Omelyaneko, VG. 2, Knutson, K1. 1University of Utah; 2Watson Laboratories, UT.

Enhanced Intestinal Absorption of P-GP Substrates by Apricot Extract. Deferme, S., Augustijns, P. Catholic University of Leuven, Belgium

Turnover and Transport Dynamics of Glutathione in Modulating Ocular Surface Health. Gukasyan, HJ., Kannan, R., Kim, K., Trousdale, MD., Lee, VH. University of Southern California, CA

Pore Controlled Diffusion of Nanospheres Across Bovine Nasal Mucosa Limits its Suitability to Assess Intracellular Translocation. Koch, A., Ziegler, U., Merkle, HP. Swiss Federal Institute of Technology, Zürich, Switzerland

Genechip Expression Analysis and Correlation to Valacyclovir Pharmacokinetics in Healthy Volunteers. Landowski, CP., Sun, D., Menon, SS., Ramachandran, C., Barnett, JL., Foster, DR., Welage, LS., Amidon, GL. The University of Michigan, MI

The Contribution of the Lymphatics to the Subcutaneous Bioavailability of Epoetin Alpha in Sheep. McLennan, DN., Porter, CJH., Charman, SA. Victorian College of Pharmacy, Monash University, Australia

An Investigation of Caffeine Buccal Permeability. Nicolazzo, JA., Reed, BL., Finnin, BC. Victorian College of Pharmacy, Monash University, Australia

The Jejunal Permeability(Peff) and Presystemic Metabolism of Isotretinoin in Humans. Petri, N., Tannergren, C., Knutson, L., Wiegand, U. Lennernäs, H. Uppsala University, Sweden

POSTER PRESENTATIONS

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Transport Characteristics of Fexofenadine in the Caco-2 Cell Model Compared to In Vivo Permeability. Petri, N. Tannergren, C., Knutson, L., Lennernäs, H. Uppsala University, Sweden

Intestinal Absorption and Metabolism of Sulforaphane and Quercetin as well as Regulation of Phase 2 Enzymes in Human Jejunum In Vivo and in Caco-2 Cells. Petri, N., Tannergren, C., Bennett, RN., Holst, B., Bao, Y., Plumb, GW., Bacon, J., O’Leary, K., Knutson, L., Forsell, P., Eriksson, T., Lennernäs, H., Williamson, G. Uppsala University, Sweden

Effect of Liquid Volume and Food Intake on Danazol Bioavailability. Sunesen, R., Vedelsdal, R., Kristensen, HG., Christrup, L., Müllertz. The Royal Danish School of Pharmacy, Denmark

A Quantitative Study of AC Residual Effect with HEM. Yan, G., Li, SK., Zhu, H., Higuchi, WI. University of Utah, UT

Analytical Methods

Investigation of Substance P Metabolism using Capillary Electrophoresis with Laser-Induced Fluorescence and Liquid Chromatography with Tandem Mass Spectrometry. Cooper, JD., Freed, AL., Audus, KA., Lunte, SM. The University of Kansas, KS

Analysis of Peptides and Peptide-Based Pharmaceuticals using Microchip Electrophoresis Systems. Lacher, NA., Vandaveer, IV., Martin, RS., Lunte, SM. The University of Kansas, KS

Assay for the Determination of Homocysteine as a Marker for Cardiovascular Disease. Pasas, SA., Davies, MI., Lunte, SM. The University of Kansas, KS

POSTER PRESENTATIONS

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Use of Microdialysis to Evaluate Dexamethasone Tissue Distribution after Oral and Topical Administration. Schuck, V., Muller, M., Grant, M., Hochhaus, G., Hartmut, D. University of Florida, FL

Isoperibol Calorimetry Studies of Protein/Amino Acid Interactions in Solid State. Tian, F., Rytting, JH. The University of Kansas, KS

Pharmacokinetics

A Novel Indirect Response PK-PD Model to Characterize the Effect of Faropenem on Streptococcus Pneumoniae and Haemophilus Influenzae. Khunvichai, A., Hirt, M., Karmann, M., Rand, K., Derendorf, H.

Predictions of Dose-Dependent Pharmacokinetics of Prednisolone. Kurz, J., Bhattaram, VA., Derendorf, H. University of Florida, FL.

Pharmacokinetics and Pharmacodynamics of Cefpodoxime and Cefixime. Liu, P. 1, Müller, M. 2, Grant, M. 1, Rand,K. 1, Derendorf, H1. 1University of Florida, FL; 2Vienna University School of Medicine

In Vitro Pharmacodynamic Modeling of Ceftriaxone. Liu, Q., Lamberty, N., Hintze, S., Rand, K., Derendorf, H. University of Florida, FL

Physical Characterization

The Effect of Solution Environment on Second Virial Coefficient of IgG and its Relation to Solubility of IgG. Bajaj, H., Sharma, VK., Kalonia, DS. University of Connecticut, CT

POSTER PRESENTATIONS

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Stability of a Synthetic Human Chorionic Gonadotropin (HCG) Antigen in PLGA Microspheres. Cui, C., Schwendeman, SP. The University of Michigan, MI

Microclimate pH Inside PLGA Films and a Possible Relationship with Water-Soluble Acids. Ding, AG., Schwendeman, SP. The University of Michigan, MI

Preformulation/Formulation Devlopment of a Novel Interferon. Katayama, DS. 1, Villarete, L. 2, Liu, CP. , Nayar, R. 3, Manning, MC1,3. 1University of Colorado Health Sciences Center; 2Pepgen Corporation; 3HTD Biosystems

Feasibility Studies of Contact Lenses as a Potential Tear Collection Device. Goteti,K., Kern, S., Andrade, J. University of Utah, UT

The Effect of Water on the Solubility of Steroid Drugs in Medium and Long Chain Triglycerides. Land, LM., Bummer, PM. University of Kentucky, KY

Effect of Formaldehyde Treatment on the Inactivation of Diphtheria Toxin. Metz, B. 1,2, Jiskoot, W. 1, Hennink, W. 1, Crommelin, D1., Kersten, G2. Utrecht University; National Institute of Public Health and the Environment, The Netherlands

Metal Catalyzed Oxidation of Insulin. Sadineni, V., Schöneich, C. The University of Kansas, KS

Residual Polyvinyl Alcohol Associated with Poly(D,L-Lactide-Co-Glycolide) Nanoparticles Affects their Physical Properties and Cellular Uptake. Sahoo, SK., Panyam,J., Prabha, S., Labhasetwar, V. University of Nebraska Medical Center, NE

POSTER PRESENTATIONS

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Stabilization of Proteins by Amorphous Mannitol During Vacuum Drying of [Beta]-Lactoglobulin

Following Precipitation by PEG. Sharma, VK., Kalonia, DS. University of Connecticut, CT

Preparation of Solid Dispersions of the Class II drug Itraconazole with Hydrophilic Polymers Prepared by Hot-Stage Extrusion. Six, K. 1, Leuner, C. 2, Dressman, J. 2, Van den Mooter, G1. 1University of Leuven, Belgium; 2University of Frankfurt, Germany

Kinetics of Asparagine Deamidation in a (-Hairpin Peptide. Stotz, CE., Vander Velde, D., Topp, EM. The University of Kansas, KS.

Studying the Reaction Between Enantiomers in Solid State Using X-Ray Powder Diffraction. Tian, J. 1, Borchardt, TB. 2, Stella, VJ1. 1The University of Kansas, KS; 2Pharmacia Corp, IL

Modeling

Human Nasal Cell Culture Model to Screen Subacute Effects of Pharmaceutical Excipients on Ciliary Activity. Agu, RU., Jorissen, M., Willems, T., Kinget, R., Verbeke, N. K.U Leuven, Belgium

Modeling Multi-Drug Interactions Using a Modified MWC Model. Fidler, ML, Egan, TD., Kern, SE. University of Utah, UT

In Silico Metabolism: Semi-Empirical Calculation of Hydrogen-Atom Abstraction Energy for 42 Common Drugs as a Model of P450 Oxidation. Wang, J., Bolger, MB., University of Southern California, CA

Podia Presentations

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Podia 01-14: Wednseday, November 6, 2002

Podia 15-27: Thursday, November 7, 2002

Name Podia Number

Meagan Anderson………………………………07

Vikram Arya…………………………………… .01

KareemAzab……………………………………..08

Corbin Bachmeier………………………………. 25

Maytal Bivas-Benita ..…………………………...13

R. Tyler DeGraw………………………………....03

Annelies Derycke………………………………...15

Hui Ding…………………………....…………….16

Carsten Ehrhardt………………………………….04

Andre Eriksson………………...…….…………...26

Maki Hasegawa………...…….…………………. 23

Akhiro Inano……………………………………...27

Samantha Jilek………………………………........11

Jennifer Links…………………………………….24

Zak Megeed……………………………....………22

Erem Memisoglu…………………………………17

Thomas Merdan…………………………………..20

Sarita Naik………………………………………..18

Benjamin Ross……………………………………14

Alicia Segrave ……………………………………02

Lena Strindelius……………....…………………..12

Marc Sutter…………...….………………………..05

Charlie Tang…...…….……………………………19

Karin Wensink….……………...………………….06

Karen White……………………………………….10

Kei Yasuda………....……………………………...21

Huiyu Zhou………….………………...…………..09

Podium 01

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DEVELOPMENT OF A BOOTSTRAP PROCEDURE FOR RESAMPLING PHARMACOKINETIC DATA

V. Arya1, A. Agarwal2, Elziria Nunan3, Tasso Santos3, G. Hochhaus1

1Department of Pharmaceutics , 2 Department of Decision and Information Sciences, University of Florida, Gainesville, FL & 3 Universidade Federal De Minas Gerias, Brazil

Purpose. To develop a general and user friendly bootstrap application for resampling pharmacokinetic data when complete concentration time profiles from individual animals cannot be obtained.

Methods. The bootstrap application was developed using visual basic in Excel. The application was divided into four sections: user form, input data sheet, output sheet, and the statistical analysis sheet. The user form allowed the entry of information such as dose, number of time points, animals at each time point etc. The input data sheet was used to enter plasma concentration values available at each time point. After the data input, the program was designed to randomly select one concentration value from the pool of concentration values at a given time point. This was iterated for all the other time points to generate resampled concentration time profiles. Noncompartmental analysis were applied to each of the resampled concentration time profiles to compute pharmacokinetic parameters like Cmax, Tmax, AUC, AUMC, MRT, KE, and T1/2. These parameters were tabulated in the output sheet. Statistical comparisons (t test and ANOVA) of the parameters was done in the statistical analysis sheet. The program was applied to perform a pharmacokinetic and statistical analysis of data obtained after subcutaneous administration of tytyustoxin to a group of adult and weanling rats.

Results. The application used the resampled concentration time profiles to calculate and statistically compare the pharmacokinetic parameters for the adult and weanling groups. All the calculated pharmacokinetic parameters were statistically significantly different (p PLtoB). Probenecid lowered ddI PBtoL but did not affect PLtoB. F-ddA metabolism was similar in both directions, with metabolite back-flux accounting for most of the F-ddA absorbed.

Conclusions. This technique is a powerful tool to examine intestinal wall metabolism and transport. F-ddA, F-ddI, and ddI showed asymmetric transport, with higher than expected blood to lumen flux. ddI blood to lumen flux is probenecid sensitive.

This work was supported by an AFPE pre-doctoral fellowship and NIH RO1 NS39178.

Podium 04

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HUMAN ALVEOLAR EPITHELIAL CELL MONOLAYERS IN PRIMARY CULTURE AS AN IN VITRO MODEL FOR DRUG ABSORPTION STUDIES

Ehrhardt C, Schaefer UF and Lehr CM.

Saarland University, Saarbruecken, Germany.

Purpose. To further develop and characterize a cell culture model for in vitro studies of pulmonary drug absorption and transport based on human alveolar epithelial cells (HAEpC) with regards to active drug transport mechanisms.

Methods. Human type II pneumocytes were cultured on filter inserts until reaching confluency. This was monitored by measuring the transepithelial electrical resistance (TEER) and the permeation of the paracellularly transported fluorescein sodium (Flu-Na). Expression of P-glycoprotein (P-gp) and lung resistance-related protein (LRP) was investigated by immunocytochemistry. To show P-gp activity, bi-directional transport studies were performed using the substrate marker Rhodamine 123 (Rh123).

Results. A good correlation between TEER of the monolayers and flux of Flu-Na could be observed with a cut off at a TEER of ~ 500 Ohms·cm2. HAEpC stained positively for P-gp and LRP. The flux of Rh123 showed a significant increase in the secretory compared to the absorptive direction.

Conclusions. HAEpC provide an good in vitro model to study drug transport and absorption in the alveolar region of the lung. The type I-like monolayers are ready for transport experiments after 8 days in culture. P-gp and LRP are expressed and show functionality.

Podium 05

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MEMBRANE PROPERTIES AFFECTING DRUG PERMEABILITY

Marc Sutter, Tamara Fiechter, Georgios Imanidis

University of Basel, Switzerland

Purpose. The relevance of order and rotational motion measured with fluorescent probe molecules embedded in phospholipid bilayers for describing the permeability properties of the membranes was studied.

Methods. Order parameters (S) and rotational correlation times (() for 1,6-diphenyl-1,3,5-hexatriene (DPH) and for 1-(4-trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene (TMA-DPH) in unilamellar vesicles were determined by time resolved fluorescence spectroscopy. Liposomes consisted of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine (POPC) and 0 to 30 mole-% cholesterol (chol). Permeability coefficients (P) were measured at 25°C using [14C]-mannitol.

Results. P decreased from 3.6 to 1.5 *10-11cm/s with rising chol content while S increased from 0.37 to 0.79 (DPH) and from 0.60 to 0.84 (TMA-DPH). There was a negative correlation between S and P in both cases. No clear dependence of ( on chol content was observed. ( showed a greater variation than S especially for DPH.

Conclusions. S reflects the order of the phospholipid hydrocarbon chains over a considerable length of the chains. Therefore membrane order is clearly an important parameter controlling permeability. Differences in S for DPH and for TMA-DPH may be caused by the different location of these probes. Greater changes in ( may be reached in other membrane systems or by temperature variation.

Podium 06

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Quantitative release of active lysozyme from dextran based hydrogels

Wensink K.D.F1, Jiskoot W.1, Verrijk R.2, Hennink W.E.1

1Department of Pharmaceutics, Utrecht Institute for Pharmaceutical Sciences (UIPS), Utrecht University;2Research department, Octoplus Technologies b.v., Leiden

Purpose: To investigate whether a model protein (lysozyme) was released quantitatively and in its intact form from dextran based hydrogels.

Methods: Lysozyme loaded dex-HEMA hydrogels varying in water content (50, 70, 90 %) and in degree of methacrylate substitution (5, 7.7, 15) were prepared. All gels were incubated in PBS at 37 oC. Once the gels were fully degraded, total protein concentration was determined with the BCA protein assay. Moreover, the enzymatic activity assay of the released protein was determined, the structural integrity with SDS-PAGE (aggregation, fragmentation) and HPLC and mass spectrometry (oxidation). Finally, insoluble hydrogel fragments were analyzed with FTIR for adsorbed lysozyme.

Results: For all gels total lysozyme recovery was nearly quantitative. The specific activity of the released lysozyme was substantially maintained. SDS-PAGE indicated no fragmentation or aggregation. HPLC and mass spectrometry showed that part of the released lysozyme was oxidized. Inspection with FTIR of the insoluble material did not show the presence of lysozyme.

Conclusions: Nearly quantitative release of lysozyme from dex-HEMA hydrogels was obtained with preservation of its structural integrity and functional activity.

Podium 07

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Selectivity of MTX-ICAM-1-peptide conjugate compared to MTX

Anderson, ME, Yakovleva T, Jean-Farve DJ, and Siahaan TJ.

University of Kansas, Lawrence KS

Purpose. The objective of this work is to study the activity of MTX-ICAM-1 peptide conjugates to deliver MTX to activated T-cells that express LFA-1 receptors. We have developed an MTX-ICAM-1-peptide conjugate that can bind to and is internalised by activated T-cells via LFA-1.

Methods. 96-well plates were coated with soluble LFA-1 in order to determine the relative affinity of these peptides for LFA-1 and their ability to block anti-LFA-1 antibody binding. A Dojindo cytoxicity kit was used to compare the activity between MTX-ICAM-1-peptide conjugates and MTX in MOLT-3 T-cells. As a control, an MTT assay was used to establish the activity of MTX-peptides in KB cells, which do not express LFA-1. TNF-( production by leukocytes after treatment with ICAM-1 peptides, MTX-ICAM-1 peptide conjugates, and MTX were evaluated using ELISA.

Results. We demonstrated that peptides derived from ICAM-1, bind LFA-1 coated plates in a specific manner and were capable of inhibiting anti-LFA-1 antibodies. We found the activity of MTX-peptide conjugates in the KB cell line to be minimal. We found that the MTX-peptide conjugates were more potent than MTX in activated T-cells than in resting T-cells. The ability of MTX-peptide conjugates to inhibit TNF-( production in activated T-cells was very similar compared with MTX.

Conclusions. We found MTX-peptide conjugates only active in cells that have LFA-1 receptor. Also these MTX-peptide conjugates were more effective in activated T-cells compared to non-activated T-cells. The unique binding and internalisation characteristics of these MTX-peptide conjugates may be utilised to reduce the side effects of MTX in treating autoimmune diseases.

Podium 08

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NEW BORONATED, CATIONIZED ACRYLAMIDE CO-POLYMERS: SYNTHESIS AND MUCOSAL ATTACHMENT PROPERTIES

Kareem Azab, Morris Srebnik and Abraham Rubinstein

The Hebrew University of Jerusalem, Israel.

Purpose. (1) To synthesize novel boronated cationized acryl amide copolymers for the local treatment of colon cancer in boron neutron capture therapy; (2) to quantify the effect of cationization of the polymers on their adherence to the cecum of the rat in vitro.

Methods. The acryl amide monomers was prepared by protecting one amine group of 1,2-diaminoethane with the BOC group, and the hydroxyl groups of 3-aminophenylboronic acid with pinacol, followed by reacting the free amine groups with acryloyl chloride. Four different copolymers were prepared, containing the following ratios of the cationized monomer: 30, 50, 80 and 100 mole%. After polymerization the BOC and the pinacol protecting groups were removed. Mucoadhesion was assessed by a self-assembled detachment apparatus, which measures the detachment force between each polymer solution and the cecal epithelium.

Results. The detachment force was directly proportional to the relative amount of cationized monomers in the copolymer. The detachment force values were 2.4 ±0.24, 2.3±0.66, 3.5±0.34, 3.7±0.68 and 4.6±0.28 g-force for the blank, 30, 50, 80 and 100 mole% of the cationized products respectively.

Conclusion. Due to their mucosal attachment properties cationized boronated acrylamide copolymers are potential platforms for local drug delivery in the colon.

The Study was supported by a grant # 663/99-2 from the Israeli Science Foundation

Podium 09

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Incorporation of Isoniazid into Poly(L-lactide) Microspheres Using a Prodrug Approach

H. Zhou, D. L. Biggs, D. J. Claffey, J. A. Ruth, T. W. Randolph, K. Ng, M. C. Manning.

University of Colorado Health Sciences Center, Denver, CO

Purpose. The use of precipitation with a compressed antisolvent (PCA), combined with hydrophobic ion pairing (HIP), to manufacture poly(lactide) microspheres containing ionic therapeutic agents has been demonstrated previously. Using a prodrug approach followed by HIP-PCA, a neutral anti-tuberculosis drug, isoniazid (isonicotinic acid hydrazide, INH), was incorporated into poly(L-lactide) (PLA) microparticles for pulmonary delivery.

Methods. A sodium salt of isoniazid methanesulfonate (Na-INHMS) was synthesized using a standard chemical synthesis method. Isoniazid methanesulfonate (INHMS) was ion-paired with either tetrapentylammonium or tetraheptylammonium in dichloromethane/water partition experiments. The resultant HIP complexes were extracted into the organic phase, purified with flash chromatography and incorporated into PLA microspheres by precipitation with supercritical carbon dioxide. Particle size and morphology were determined using scanning electron microscopy (SEM). Release studies were performed in phosphate-buffered saline (PBS) (pH 5.0 or 7.4). The drug concentration in the receiver medium was measured by HPLC.

Results. The logarithm of the dichloromethane/water partition coefficient (log P) was 0.304 for tetrapentylammonium-INHMS and 1.43 for tetraheptylammonium-INHMS, compared to a value of –1.80 for Na-INHMS. The significant increases in log P suggested enhanced solubility of INHMS in organic solvents, allowing subsequent incorporation of the drug in PLA microspheres using PCA. The microspheres have a mean size of 2 microns with a standard deviation of 1.37 microns. Drug loading of these microspheres was 7.6(0.6% for tetrapentylammonium-INHMS and 5.6(0.3% for tetraheptylammonium-INHMS. Microspheres containing tetrapentylammonium-INHMS complexes exhibited an initial burst of ~65% and those containing tetraheptylammonium-INHMS complexes displayed an initial burst of ~60%. After the first few hours the cumulative drug release was linear with the square root of time for both types of microspheres. The pH of the receiver medium does not seem to affect the release kinetics. The drug can be detected in the receiver medium for up to 30 days.

Conclusions. A prodrug approach can be employed to introduce a charged moiety into an otherwise neutral molecule, allowing subsequent microencapsulation of the molecule into PLA using the HIP-PCA approach. Despite the relatively large initial bursts, the drug-loaded microspheres afterwards exhibited release profiles consistent with a matrix-controlled release mechanism.

Podium 10

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LIPOSOMAL DELIVERY OF ANTIGEN TO HUMAN DENDRITIC CELLS

KL White, MJ Copland, MA Baird, S Hook, NM Davies, T Rades.

University of Otago, Dunedin, New Zealand

Purpose. To determine whether uptake of liposomally entrapped antigen enhances activation of human dendritic cells and whether this results in a corresponding increase in the ability of these cells to induce proliferation and activation of primed T cells.

Methods. Liposomes composed of varying concentrations of mannosylated lipids, containing FITC-labelled ovalbumin or tetanus toxoid were prepared by hydration of a lipid film with an aqueous solution of protein. Immature human dendritic cells (DC) were generated from human peripheral blood monocytes by culture with GM-CSF and IL-4. Expression of cell surface activation markers by DC was investigated by fluorescence flow cytometry. T cell proliferation and cytokine production was measured before and after activation by fluorescence flow cytometry.

Results. Exposure to liposome entrapped antigen resulted in enhanced expression of cell surface markers indicative of DC activation and maturation compared to antigen in solution. These phenotypic changes were reflected in the enhanced ability of liposomal tetanus toxoid exposed DC to stimulate proliferation of primed T cells.

Conclusions. Liposome entrapped antigen leads to enhanced DC activation and subsequent T cell proliferation compared to antigen in solution.

Podium 11

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PHAGOCYTOSIS OF BIODEGRADABLE

MICROPARTICLES TO INDUCE MATURATION AND ACTIVATION OF DENDRITIC CELLS

Samantha Jilek, Hans P. Merkle and Elke Walter

Institute of Pharmaceutical Sciences, ETH Zürich, 8057 Zürich, Switzerland

Purpose. The focus of this project was to study the capacity of biodegradable microparticles (MS) to induce maturation and activation of dendritic cells (DC).

Methods. MS were prepared by spray drying using biodegradable polymers, poly(D,L-lactide) (PLA) and poly(D,L-lactide-co-glycolide) (PLGA). Plasmid DNA or salmon DNA were either encapsulated, coated onto MS or added to MS in solution. MS carried negative charges. Positively charged MS were obtained by co-encapsulation of polyethylenimine. Human blood monocyte-derived DC were differentiated over 7 days before incubation with MS. On day 10, the presence of maturation and activation surface markers (CD83 and CD86, respectively) was assessed by flow cytometry.

Results. Parallel maturation (CD83) and activation (CD86) of DC were induced by LPS only. Upon phagocytosis, PLGA MS induced either upregulation of CD83 (negative charge) or CD86 (positive charge). PLA MS failed to induce upregulation of CD83 or CD86. In addition to CD83, PLGA MS-matured DC produce IL-12, a marker for Th1 response, and TNF-(, requested for recruitment and maturation/activation of immune cells at the site of inflammation.

Conclusions. Our results suggest that the character of the phagocytosed microparticles determines the response of DC in terms of maturation or activation and cytokine production.

Podium 12

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FLAGELLIN FROM SALMONELLA ENTERITIDIS INDUCE STRONG IMMUNE RESPONSES AFTER ORAL AND NASAL VACCINATION IN C3H/HeJ MICE

Lena C. Strindelius and Ingvar Sjöholm.

Dept. of Pharmacy, Uppsala University, BMC, Box 580, SE-751 23 Uppsala, Sweden

Purpose. The aim of this study was to investigate the immune response elicited in C3H/HeJ mice after immunization with flagellin (FliC) from Salmonella enteritidis, conjugated to poly(acrylstarch) microparticles, or co-administered with rCTB as an adjuvant.

Methods. Different groups of C3H/HeJ mice were immunized orally or nasally on three consecutive days or subcutaneously. A booster was given 21 days after priming. Blood and fecal samples were collected and analyzed with ELISA for total IgG and IgM and subclasses in serum, and IgA in feces. The degree of protection was studied by challenging the mice orally with Salmonella enteritidis.

Results. The humoral response showed that all groups induced high FliC-antibody titers, especially the nasally immunized group and the group that received antigen conjugated microparticles orally. The mucosal immune response showed that the nasally immunized group had the highest amount of IgA in feces. The subclass profile indicated that a Th1 response was induced.

Conclusion. This study shows that an impressive immune response can be induced by FliC, even when administered orally or nasally.

Podium 13

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PULMONARY DNA VACCINATION AGAINST TB USING CHITOSAN NANOPARTICLES

Bivas-Benita, M., Geluk, A., van Meijgaarden, K. E., van der Borch, F., Ottenhoff, T.H.M., Junginger, H. E., Borchard, G.

Leiden University, Leiden, The Netherlands

Purpose. Determination of the immunogenicity of a Mycobacterium tuberculosis (M. TB) DNA vaccine, given as an aerosol to murine lungs using chitosan nanoparticles.

Methods. Chitosan nanoparticles were made according to a complexation-coacervation method. In-vivo vaccination was done with HLA-A2 transgenic mice using a plasmid encoding a poly-epitope protein containing various M. TB-derived T cell epitopes. Mice were given 3 DNA doses followed by a protein boost. Endotracheal application was done using a Penn-Century microsprayer and i.m. injection in the hind leg used for control. Evaluation of immunization efficiency was done by proliferation and IFN-( secretion assays, using M. TB and BCG as antigens.

Results. The endotracheal application showed equal and in some cases superior results compared to i.m. injection for both parameters measured. Enhanced T cell proliferation and IFN-( secretion by splenocytes was seen when chitosan nanoparticles were used.

Conclusions. The endotracheal administration of this DNA vaccine seems more efficient compared to i.m. immunization. Pulmonary vaccination with chitosan nanoparticles increases the T cell response and therefore represents a potential non-viral delivery system for TB DNA vaccines.

Podium 14

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SYNTHESIS AND IMMUNOLOGICAL EVALUATION OF VACCINES CONTAINING MULTIPLE COPIES OF PEPTIDE EPITOPES

Ross BP1, McGeary RP1, Horváth A1, Olive C2, Good M2 and Toth I1.

1School of Pharmacy, The University of Queensland, Brisbane, Queensland, Australia; 2CRC for Vaccine Technology, QIMR, Brisbane, Queensland, Australia.

Purpose. To examine the immunogenicity of synthetic vaccines containing multiple copies of Group A Streptococcus (GAS) conserved and/or variable epitope(s) on a lipophilic multiplying core.

Methods. Synthetic vaccines were constructed by solid phase peptide synthesis. These constructs consisted of multiple copies of GAS peptide epitopes, as well as several lipoamino acids which act as an internal carrier-adjuvant system. B10.BR mice were given primary immunisation on day 0, and then boosted on days 21, 28 and 35. Antibody titres were determined by ELISA.

Results. The synthetic vaccines were more immunogenic than the peptides alone. For example, at day 14, antibodies against the construct containing 4 copies of the peptide LRRDLDASREAKKQVEKALE were detected whereas no antibodies were detected against the non-conjugated peptide. In another case, titres were up to 3000 times greater for the conjugated peptide than for the non-conjugated peptide.

Conclusions. Synthetic vaccines incorporating multiple copies of the epitopes and lipoamino acids were significantly more immunogenic than the non-conjugated peptides with conventional adjuvants. Such synthetic vaccines show potential for immunisation.

Podium 15

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TRANSFERRIN-MEDIATED TARGETING OF AlPcS4 ENCAPSULATED IN STERICALLY STABILIZED LIPOSOMES

AS Derycke, PA de Witte.

KULeuven, Belgium

Purpose. To increase the tumorselectivity and photodynamic efficacy of the photosensitizer AlPcS4 by encapsulation in transferrin-targeted sterically stabilized liposomes (Tf-Lip-AlPcS4).

Methods. The cellular accumulation of Tf-Lip-AlPcS4 was determined by incubating AY27 cells for 4h or 24h with a 10µM solution, following MeOH extraction and fluorescence measurements. In similar experiments the receptor-mediated endocytosis was inhibited by colchicine or cytochalasin B. The photodynamic efficacy was investigated by incubating AY27 cells for 24h with a 10µM solution and subsequent irradiation (651nm, 4.5J/cm2). The photocytotoxicity was determined by the antiproliferative MTT assay. Analogues experiments were performed using non-targeted liposomal and free AlPcS4.

Results. The intracellular accumulation of Tf-Lip-AlPcS4 exceeded that of non-targeted liposomal and free AlPcS4 with a factor 100 respectively 10. Inhibition of receptor-mediated endocytosis completely inhibited the intracellular uptake. Tf-Lip-AlPcS4 and free AlPcS4 showed similar photocytotoxicity, while no effect was observed in the case of the non-targeted liposomes.

Conclusions. Transferrin-targeted sterically stabilized liposomes are efficient tools for improving the tumor-selectivity and efficacy of AlPcS4. Intracellular uptake occurs via receptor-mediated endocytosis.

Podium 16

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DESIGN OF A Lymphoma TARGETED DRUG DELIVERY SYSTEM: Selection of CD21 recognizable Epitopes with Phage Display

H. Dinga, W.M. Prodingerb, J. Kopečeka

aUniversity of Utah, Salt Lake City UT

bUniversity of Innsbruck, Austria

Purpose. To identify CD21 receptor-binding epitopes using phage display libraries. To use selected epitopes as targeting moieties in the design of a lymphoma targeted drug delivery system.

Methods. Phage libraries, the fUSE5 15-mer library and the M13KE 10-mer library, were used for the selection of epitopes recognizable by CD21 receptors. For each phage display, four rounds of selections were performed. The affinity of selected phages was verified with phage ELISA. The gold surface of Spreeta biosensor was modified with NTA-SH. His-tagged truncated CD21 receptor was then immobilized through Ni-His tag chelation. The affinity of a synthesized peptide containing selected sequence was analyzed with surface plasmon resonance (SPR) using a modified Spreeta biosensor.

Results. Both phage selections resulted in consensus sequences-ARVPFRFQLNSSGPV and HFATRSLLLG. Both phage ELISA and preliminary SPR experiments indicated a specific binding of the selected sequence ARVPFRFQLNSSGPV to the CD21 receptor.

Conclusions. The selected peptide sequences are good candidates as targeting moieties. In future experiments, they will be used in the design of HPMA copolymer based systems biorecognizable by lymphomas.

Podium 17

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PREPARATION AND CHARACTERIZATION OF PROGESTERONE-LOADED AMPHIPHILIC ß-CYCLODEXTRIN NANOSPHERES

Erem Memişoğlu, Amelie Bochot, Dominique Duchene and A. Atilla Hincal

Hacettepe University, Ankara TURKEY

Université Paris-Sud, Chatenay-Malabry FRANCE

Purpose. To formulate and characterize nanospheres prepared directly from progesterone-amphiphilic ß-CD inclusion complexes without the aid of surfactants.

Methods. Progesterone was incorporated intoamphiphilic ß-CD molecule (1:2 rug:amphiphilic ß-CDmolar ratio) and the formation of a genuine complex was demonstrated by various techniques including HNMR at 400 MHz, DSC, FAB MS and FT IR spectroscopy. Nanospheres were prepared directly from these inclusion complexes using the nanoprecipitation technique and further characterized by particle size analysis by dynamic light scattering, zeta potential measurement and freeze-fracture microscopy. Entrapped drug content in the nanospheres were determined by HPLC. Finally, the release of progesterone into water:PEG400 (60:40) release medium was evaluated in order to fully characterize amphiphilic ß-CD nanospheres loaded with progesterone for intravenous administration.

Results. Preparation technique enhanced the entrapment efficiency of nanospheres significantly (p ................
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